Correcting Autoinflammation in STING-Associated Vasculopathy with Onset in Infancy (SAVI) by Human Stem Cell Genome-Editing

Correcting Autoinflammation in STING-Associated Vasculopathy with Onset in Infancy (SAVI) by Human Stem Cell Genome-Editing | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Correcting Autoinflammation in STING-Associated Vasculopathy with Onset in Infancy (SAVI) by Human Stem Cell Genome-Editing Mara Pavel-Dinu, Sebastien Viel, Sridhar Selvaraj, Marc Gastou, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5090716/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Type I interferonopathies comprise a large group of Mendelian autoinflammatory diseases that generally lack effective treatments. STING-associated Vasculitis with onset in Infancy (SAVI) is a severe autosomal dominant disease due to gain-of-functon(GOF) mutations in the gene STING1, encoding for the Stimulator of Interferon Response CGAMP Interactor 1 (STING1). SAVI leads to a severe multisystemic disease comprising systemic vasculitis and lung fibrosis. A possible therapeutic approach is to engineer the patient’s hematopoietic system to restore interferon homeostasis, thereby preventing chronic inflammation and organ damage and halting disease development. Here, we describe a “universal” genome editing correction strategy using CRISPR/Cas9 that targets the stimulator of interferon response cGAMP interactor 1 (STING1) gene at the endogenous locus in SAVI patient-derived induced pluripotent stem cells (iPSCs) and in human CD34+ hematopoietic stem and progenitor cells (HSPCs). The engineered SAVI-iPSCs express normal levels of STING1 protein following differentiation into monocytes and macrophages and no longer produce interferon-alpha (IFN-α) at a basal state. Using SAVI-gene-engineered HSPCs, we determined the minimum fraction of mutant alleles required to induce spontaneous IFN-α production, thus establishing the threshold of genome correction necessary to rescue the disease. Finally, we demonstrated engineered CD34+ HSPCs at the STING1 locus retain regenerative potential by supporting long-term repopulating capacity and multi-lineage differentiation potential following transplantation into immunocompromised mice. Together, these studies established the rationale for the clinical translation of SAVI genome editing and a therapeutic framework for correcting other type I interferonopathies. Biological sciences/Stem cells/Haematopoietic stem cells Biological sciences/Immunology/Autoimmunity Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Full Text Additional Declarations There is NO Competing Interest. Fibroblasts were cultured from a skin biopsy specimen obtained from a SAVI patient upon informed consent according to the IRB protocol 00013204 reviewed and approved by the Scientific and Ethics Committee of the Civil Hospices of Lyon. IRB protocol titled: CRISPAVI: Towards the development of targeted gene therapy for SAVI syndrome. All experiments using non-human vertebrates (mice) were performed per National Institutes of Health institutional guidelines and were approved by the Stanford University Administrative Pavel on Laboratory Animal Care (IACUC Protocol ID: 20565) Table 1 is available in the Supplementary Files section Supplementary Files VielSetalTable1.ai VielSetalSupplementaryFigure1.ai Dataset 1 VielSetalSupplementaryFigure2.ai Dataset 2 VielSetalSupplementaryFigure3.ai Dataset 3 VIelSetalSupplementaryFigure4.ai Dataset 4 VielSetalSupplementaryFigure5.ai Dataset 5 VielSetalSupplementaryTable1.xlsx Dataset 6 ReportingSummary.pdf Reporting Summary Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5090716","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":355341820,"identity":"2d335d59-860e-469b-9697-1d53f1c07699","order_by":0,"name":"Mara 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IRB protocol titled: CRISPAVI: Towards the development of targeted gene therapy for SAVI syndrome.\u003c/p\u003e\n\u003cp\u003eAll experiments using non-human vertebrates (mice) were performed per National Institutes of Health institutional guidelines and were approved by the Stanford University Administrative Pavel on Laboratory Animal Care (IACUC Protocol ID: 20565)\u003c/p\u003e\n\u003cp\u003eTable 1 is available in the Supplementary Files section\u003c/p\u003e","formattedTitle":"Correcting Autoinflammation in STING-Associated Vasculopathy with Onset in Infancy (SAVI) by Human Stem Cell Genome-Editing","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-5090716/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5090716/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Type I interferonopathies comprise a large group of Mendelian autoinflammatory diseases that generally lack effective treatments. STING-associated Vasculitis with onset in Infancy (SAVI) is a severe autosomal dominant disease due to gain-of-functon(GOF) mutations in the gene STING1, encoding for the Stimulator of Interferon Response CGAMP Interactor 1 (STING1). SAVI leads to a severe multisystemic disease comprising systemic vasculitis and lung fibrosis. A possible therapeutic approach is to engineer the patient’s hematopoietic system to restore interferon homeostasis, thereby preventing chronic inflammation and organ damage and halting disease development. Here, we describe a “universal” genome editing correction strategy using CRISPR/Cas9 that targets the stimulator of interferon response cGAMP interactor 1 (STING1) gene at the endogenous locus in SAVI patient-derived induced pluripotent stem cells (iPSCs) and in human CD34+ hematopoietic stem and progenitor cells (HSPCs). The engineered SAVI-iPSCs express normal levels of STING1 protein following differentiation into monocytes and macrophages and no longer produce interferon-alpha (IFN-α) at a basal state. Using SAVI-gene-engineered HSPCs, we determined the minimum fraction of mutant alleles required to induce spontaneous IFN-α production, thus establishing the threshold of genome correction necessary to rescue the disease. Finally, we demonstrated engineered CD34+ HSPCs at the STING1 locus retain regenerative potential by supporting long-term repopulating capacity and multi-lineage differentiation potential following transplantation into immunocompromised mice. Together, these studies established the rationale for the clinical translation of SAVI genome editing and a therapeutic framework for correcting other type I interferonopathies.","manuscriptTitle":"Correcting Autoinflammation in STING-Associated Vasculopathy with Onset in Infancy (SAVI) by Human Stem Cell Genome-Editing","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-10-21 10:38:50","doi":"10.21203/rs.3.rs-5090716/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"e9feb0cd-955a-4dbd-a66a-c4d2d3aa4f5b","owner":[],"postedDate":"October 21st, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":37774002,"name":"Biological sciences/Stem cells/Haematopoietic stem cells"},{"id":37774003,"name":"Biological sciences/Immunology/Autoimmunity"}],"tags":[],"updatedAt":"2025-07-03T17:45:15+00:00","versionOfRecord":[],"versionCreatedAt":"2024-10-21 10:38:50","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5090716","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5090716","identity":"rs-5090716","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}